Skip to main content
NCBI home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1997 Mar;6(3):725–727. doi: 10.1002/pro.5560060323

Crystallization and preliminary X-ray diffraction studies of aSFP, a bovine seminal plasma protein with a single CUB domain architecture.

J M Dias 1, A L Carvalho 1, I Kölln 1, J J Calvete 1, E Töpfer-Petersen 1, P F Varela 1, A Romero 1, C Urbanke 1, M J Romão 1
PMCID: PMC2143676  PMID: 9070456

Abstract

Bovine acidic seminal fluid protein (aSFP) is a 1.29 kDa polypeptide of the spermadhesin family built by a single CUB domain architecture. The CUB domain is an extracellular module present in 16 functionally diverse proteins. To determine the three-dimensional structure of aSFP, the protein was crystallized at 21 degrees C by vapor diffusion in hanging drops, using ammonium sulfate, pH 4.7, and polyethyleneglycol 4,000 as precipitants, containing 10% dioxane to avoid the formation of clustered crystals. Elongated prismatic crystals with maximal size of 0.6 x 0.3 x 0.2 mm3 diffract to beyond 1.9 A resolution and belong to space group P2(1)2(1)2(1), with cell parameters a = 52.4 A, b = 41.5 A, c = 48.2 A. There is one aSFP molecule per asymmetric unit, which corresponds to a crystal volume per unit molecular mass of 2.04 A3/Da, and analytical ultracentrifugation analysis show that aSFP is a monomeric protein.

Full Text

The Full Text of this article is available as a PDF (343.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bork P., Beckmann G. The CUB domain. A widespread module in developmentally regulated proteins. J Mol Biol. 1993 May 20;231(2):539–545. doi: 10.1006/jmbi.1993.1305. [DOI] [PubMed] [Google Scholar]
  2. Calvete J. J., Dostàlova Z., Sanz L., Adermann K., Thole H. H., Töpfer-Petersen E. Mapping the heparin-binding domain of boar spermadhesins. FEBS Lett. 1996 Feb 5;379(3):207–211. doi: 10.1016/0014-5793(95)01513-2. [DOI] [PubMed] [Google Scholar]
  3. Calvete J. J., Sanz L., Dostàlovà Z., Töpfer-Petersen E. Characterization of AWN-1 glycosylated isoforms helps define the zona pellucida and serine proteinase inhibitor-binding region on boar spermadhesins. FEBS Lett. 1993 Nov 8;334(1):37–40. doi: 10.1016/0014-5793(93)81675-p. [DOI] [PubMed] [Google Scholar]
  4. Calvete J. J., Varela P. F., Sanz L., Romero A., Mann K., Töpfer-Petersen E. A procedure for the large-scale isolation of major bovine seminal plasma proteins. Protein Expr Purif. 1996 Aug;8(1):48–56. doi: 10.1006/prep.1996.0073. [DOI] [PubMed] [Google Scholar]
  5. Dostàlovà Z., Calvete J. J., Sanz L., Hettel C., Riedel D., Schöneck C., Einspanier R., Töpfer-Petersen E. Immunolocalization and quantitation of acidic seminal fluid protein (aSFP) in ejaculated, swim-up, and capacitated bull spermatozoa. Biol Chem Hoppe Seyler. 1994 Jul;375(7):457–461. doi: 10.1515/bchm3.1994.375.7.457. [DOI] [PubMed] [Google Scholar]
  6. Einspanier R., Amselgruber W., Sinowatz F., Henle T., Röpke R., Schams D. Localization and concentration of a new bioactive acetic seminal fluid protein (aSFP) in bulls (Bos taurus). J Reprod Fertil. 1993 May;98(1):241–244. doi: 10.1530/jrf.0.0980241. [DOI] [PubMed] [Google Scholar]
  7. Einspanier R., Einspanier A., Wempe F., Scheit K. H. Characterization of a new bioactive protein from bovine seminal fluid. Biochem Biophys Res Commun. 1991 Sep 16;179(2):1006–1010. doi: 10.1016/0006-291x(91)91918-3. [DOI] [PubMed] [Google Scholar]
  8. Einspanier R., Krause I., Calvete J. J., Töfper-Petersen E., Klostermeyer H., Karg H. Bovine seminal plasma aSFP: localization of disulfide bridges and detection of three different isoelectric forms. FEBS Lett. 1994 May 9;344(1):61–64. doi: 10.1016/0014-5793(94)00362-9. [DOI] [PubMed] [Google Scholar]
  9. Ensslin M., Calvete J. J., Thole H. H., Sierralta W. D., Adermann K., Sanz L., Töpfer-Petersen E. Identification by affinity chromatography of boar sperm membrane-associated proteins bound to immobilized porcine zona pellucida. Mapping of the phosphorylethanolamine-binding region of spermadhesin AWN. Biol Chem Hoppe Seyler. 1995 Dec;376(12):733–738. doi: 10.1515/bchm3.1995.376.12.733. [DOI] [PubMed] [Google Scholar]
  10. Matthews B. W. Solvent content of protein crystals. J Mol Biol. 1968 Apr 28;33(2):491–497. doi: 10.1016/0022-2836(68)90205-2. [DOI] [PubMed] [Google Scholar]
  11. Menéndez M., Gasset M., Laynez J., López-Zumel C., Usobiaga P., Töpfer-Petersen E., Calvete J. J. Analysis of the structural organization and thermal stability of two spermadhesins. Calorimetric, circular dichroic and Fourier-transform infrared spectroscopic studies. Eur J Biochem. 1995 Dec 15;234(3):887–896. doi: 10.1111/j.1432-1033.1995.887_a.x. [DOI] [PubMed] [Google Scholar]
  12. Romero A., Varela P. F., Sanz L., Töpfer-Petersen E., Calvete J. J. Crystallization and preliminary X-ray diffraction analysis of boar seminal plasma spermadhesin PSP-I/PSP-II, a heterodimer of two CUB domains. FEBS Lett. 1996 Mar 11;382(1-2):15–17. doi: 10.1016/0014-5793(96)00133-0. [DOI] [PubMed] [Google Scholar]
  13. Sanz L., Calvete J. J., Mann K., Gabius H. J., Töpfer-Petersen E. Isolation and biochemical characterization of heparin-binding proteins from boar seminal plasma: a dual role for spermadhesins in fertilization. Mol Reprod Dev. 1993 May;35(1):37–43. doi: 10.1002/mrd.1080350107. [DOI] [PubMed] [Google Scholar]
  14. Wempe F., Einspanier R., Scheit K. H. Characterization by cDNA cloning of the mRNA of a new growth factor from bovine seminal plasma: acidic seminal fluid protein. Biochem Biophys Res Commun. 1992 Feb 28;183(1):232–237. doi: 10.1016/0006-291x(92)91633-2. [DOI] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES